血管内皮生长因子D基因的原核表达及纯化

利用GST融合基因表达系统原核表达血管内皮生长因子D,并进行纯化。诱导重组质粒pGEX-VEGF-D在大肠杆菌BL21(DE3)中表达,经超声破菌后,采用GST蛋白纯化系统进行纯化,并用3C-Protease酶切去标签GST,所得产物进行15%SDS-PAGE电泳。结果表明:大肠杆菌经诱导,高效表达出分子质量约56 ku的GST-VEGF-D融合蛋白;纯化后的蛋白经酶切后电泳显示只有一条带,纯度相对较高,可基本满足生物学活性测定,为研究VEGF-D蛋白的结构和功能提供了有利的条件。

Prokaryotic Expression and Purification of Vascular Endothelial Growth Factor D

To express,purify and identify VEGF-D fusion protein,E·coli BL21(DE3) containing recombinant plasmid pGEX-6p-1-VEGF-D was induced by IPTG.The bacteria was lyzed with lysozymc.Expressed product was purified by GST protein purification system.GST was cut by 3C-protease.And then the expressed fusion protein was identified by SDS-PAGE.The result showed that the relative molecular mass of expressed product was 56 000,which was consistent with that of GST-VEGF-D fusion protein.The VEGF-D protein without GST was successfully purified.The purified protein could be used to study biological activity of VEGF-D.